Low-intensity pulsed ultrasound increases blood vessel size during fracture healing in patients with a delayed-union of the osteotomized fibula.

Disturbed vascularity leads to impaired fracture healing. Since low-intensity pulsed ultrasound (LIPUS) increases new bone formation in delayed-unions, we investigated whether LIPUS increases blood supply in delayed-unions of the osteotomized fibula, and if LIPUS-increased bone formation is correlated to increased blood supply. Blood vessel parameters were analysed using histology, immunohistochemistry, and histomorphometric analysis as well as their correlation with bone formation and resorption parameters. Fibular biopsies of thirteen patients with a delayed-union of the osteotomized fibula treated for 2-4 months with or without LIPUS originating from a randomized prospective double-blind placebo-controlled clinical trial were studied. In histological sections of the fibular biopsies parameters of blood vessel formation were measured and were related to histomorphometric bone characteristics of newly formed bone of the same samples analysed in our previously published study on the effects of LIPUS on bone healing at the tissue level in delayed-unions. LIPUS-treated delayed-unions and sham-treated delayed-unions as well as healed delayed-unions and failed-to-heal delayed-unions were compared. The volume density of blood vessels was increased in LIPUS-treated delayed-unions compared to sham-treated controls. LIPUS did not change blood vessel number, but significantly increased blood vessel size. Healed delayed-unions as well as LIPUS-treated and sham-treated delayed-unions showed significant correlations between blood vessel size and osteoid volume. LIPUS increases blood vessel size, essential for fracture healing, in bone from patients with a delayed-union of the osteotomized fibula. The increased osteoid volume in delayed-unions can largely be explained by increased blood supply and perfusion.

A histomorphometric and micro-computed tomography study of bone regeneration in the maxillary sinus comparing biphasic calcium phosphate and deproteinized cancellous bovine bone in a human split-mouth model.

OBJECTIVE: The gain of mineralized bone was compared between deproteinized bovine bone allograft (DBA) and biphasic calcium phosphate (BCP) for dental implant placement. STUDY DESIGN: Five patients with atrophic maxillae underwent bilateral sinus elevation with DBA (Bio-Oss) and BCP (Straumann BoneCeramic). After 3 to 8 months, 32 Camlog implants were placed, and biopsies were retrieved. Bone and graft volume, degree of bone mineralization, and graft degradation gradient were determined using micro-computed tomography, and bone formation and resorption parameters were measured using histomorphometry. Implant functioning and peri-implant mucosa were evaluated up to 4 years. RESULTS: Patients were prosthetically successfully restored. All but one of the implants survived, and peri-implant mucosa showed healthy appearance and stability. Bone volume, graft volume, degree of bone mineralization, and osteoclast and osteocyte numbers were similar, but BCP-grafted biopsies had relatively more osteoid than DBA-grafted biopsies. CONCLUSIONS: The BCP and DBA materials showed similar osteoconductive patterns and mineralized bone, although signs of more active bone formation and remodeling were observed in BCP- than in DBA-grafted biopsies.

Osteochondral defects of the talus: a novel animal model in the goat.

Osteochondral defects of the talus pose a difficult therapeutic challenge. An experimental animal model of the ankle joint is not available. The aim of this study was to test a newly developed animal model for osteochondral defects of the ankle in vivo. Osteochondral defects were created in the talus of goat hind legs using a posterolateral surgical approach. The defects were filled with either autologous cancellous bone or donor demineralized bone matrix or left empty as control. After 12 weeks of healing, the specimens were analyzed with radiography, macroscopy, microcomputed tomography, histology, histomorphometry, and fluorescence microscopy. It was possible to create a standardized defect in each talus. The implanted material remained in place. The analyses showed that most bony tissue was generated in the defects filled with autologous bone and least in the control defects. Our findings show that a standard osteochondral defect can be created in the talus by a relatively simple procedure in a large animal that allows qualitative and quantitative evaluation. The model can be used in future experiments to investigate alternative treatment methods before they are introduced into clinical practice.

Reaming debris as a novel source of autologous bone to enhance healing of bone defects.

Reaming debris is formed when bone defects are stabilized with an intramedullary nail, and contains viable osteoblast-like cells and growth factors, and might thus act as a natural osteoinductive scaffold. The advantage of using reaming debris over stem cells or autologous bone for healing bone defects is that no extra surgery is needed to obtain the material. To assess the clinical feasibility of using reaming debris to enhance bone healing, we investigated whether reaming debris enhances the healing rate of a bone defect in sheep tibia, compared to an empty gap. As golden standard the defect was filled with iliac crest bone. Bones treated with iliac crest bone and reaming debris showed larger callus volume, increased bone volume, and decreased cartilage volume in the fracture gap, and increased torsional toughness compared to the empty gap group at 3 weeks postoperative. In addition, bones treated with reaming debris showed increased torsional stiffness at 6 weeks postoperatively compared to the empty defect group, while bending stiffness was marginally increased. These results indicate that reaming debris could serve as an excellent alternative to iliac crest bone for speeding up the healing process in bone defects that are treated with an intramedullary nail.

Low-intensity pulsed ultrasound affects RUNX2 immunopositive osteogenic cells in delayed clinical fracture healing.

INTRODUCTION: Osteogenic cell proliferation and differentiation play an important role in adequate fracture healing, and is target for osteoinductive therapies in delayed fracture healing. The aim of this study was to investigate whether low-intensity pulsed ultrasound enhances fracture healing at the tissue level in patients with a delayed union of the osteotomized fibula through an effect on the presence of RUNX2 immunopositive osteogenic cells. The effect was studied in both atrophic and hypertrophic delayed unions. MATERIALS AND METHODS: Biopsies were obtained from 6 female and 1 male patient (age 43-63) with a delayed union of the osteotomized fibula after a high tibial osteotomy treated for 2-4 months with or without low-intensity pulsed ultrasound in a randomized prospective double-blind placebo-controlled trial. Immunolocalization of RUNX2 protein was performed to identify osteogenic cells. Histomorphometrical analysis was performed to determine the number of cells expressing RUNX2 located within and around the newly formed woven bone at the fracture end (area of new bone formation), and up to 3 mm distant from the fracture end. RESULTS: Cells expressing RUNX2 were present in all histological sections of control and low-intensity pulsed ultrasound-treated bone evaluated. Within the area of new bone formation, RUNX2 immunopositive cells were found in the undifferentiated soft connective tissue, at the bone surface (presumably osteoblasts), and within the newly formed woven bone. Low-intensity pulsed ultrasound treatment of fibula delayed unions significantly reduced the number of RUNX2 immunopositive cells within the soft connective tissue at the fracture ends, whereas the number of RUNX2 immunopositive cells at the bone surface was not affected. The number of RUNX2 immunopositive cells was similar for the atrophic and hypertrophic delayed unions. CONCLUSIONS: Immunolocalization of RUNX2 positive cells in delayed unions of the fibula reveals that delayed clinical fracture healing does not result in impairment of osteogenic cell proliferation and/or differentiation at the tissue level, even if delayed unions are clinically regarded as atrophic. Reduced number of osteogenic RUNX2 immunopositive cells within the soft connective tissue, and unchanged number of RUNX2 immunopositive cells at the bone surface, implicate that low-intensity pulsed ultrasound does not increase osteogenic cell presence, but likely affects osteogenic cell differentiation.

Low-intensity pulsed ultrasound increases bone volume, osteoid thickness and mineral apposition rate in the area of fracture healing in patients with a delayed union of the osteotomized fibula.

INTRODUCTION: Low-intensity pulsed ultrasound (LIPUS) accelerates impaired fracture healing, but the exact mechanism is unknown. The aim of this study was to investigate how LIPUS affects bone healing at the tissue level in patients with a delayed union of the osteotomized fibula, by using histology and histomorphometric analysis to determine bone formation and bone resorption parameters. MATERIALS AND METHODS: Biopsies were obtained from 13 patients (9 female, 4 male; age 42-63) with a delayed union of the osteotomized fibula after a high tibial osteotomy, treated for 2-4 months with or without LIPUS in a randomized prospective double-blind placebo-controlled trial. In the histological sections of the delayed union biopsies, 3 areas of interest were distinguished, i.e. 1) area of new bone formation at the fracture ends, 2) area of cancellous bone, and 3) area of cortical bone. Histomorphometrical analysis was performed to determine bone formation and bone resorption parameters (as well as angiogenesis). RESULTS: In LIPUS-treated delayed unions, endosteal callus formation by direct bone formation without a cartilage intermediate as well as indirect bone formation was observed, while in untreated controls only indirect bone formation was observed. In the area of new bone formation, LIPUS significantly increased osteoid thickness by 47%, mineral apposition rate by 27%, and bone volume by 33%. No increase in the number of blood vessels was seen in the newly formed bony callus. In the area of cancellous bone, bone volume was significantly increased by 17% whereas no effect on osteoid thickness and mineral apposition rate was seen. LIPUS did not affect osteoid volume, osteoid maturation time, number of osteocytes, osteocyte lacunae, or osteoclast-like cells in any of the areas of interest. CONCLUSIONS: Our results suggest that LIPUS accelerates clinical fracture healing of delayed unions of the fibula by increasing osteoid thickness, mineral apposition rate, and bone volume, indicating increased osteoblast activity, at the front of new bony callus formation. Improved stability and/or increased blood flow, but probably not increased angiogenesis, might explain the differences in ossification modes between LIPUS-treated delayed unions and untreated controls.